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An efficient Agrobacterium-mediated transformation of strawberry cv. camarosa by dual plasmid system


Citation

Haddadi, Fatemeh and Abdul Aziz, Maheran and Abdullah, Siti Nor Akmar (2015) An efficient Agrobacterium-mediated transformation of strawberry cv. camarosa by dual plasmid system. Molecules, 20 (3). pp. 3647-3666. ISSN 1420-3049

Abstract / Synopsis

An Agrobacterium-mediated transformation method was applied to introduce the luciferase reporter gene under the control of the CaMV35S promoter in the pGreen0049 binary vector into strawberry cv. Camarosa. The in vitro regeneration system of strawberry leaves to be used in the transformation was optimized using different TDZ concentrations in MS medium. TDZ at 16 µM showed the highest percentage (100%) of shoot formation and the highest mean number of shoots (24) produced per explant. Studies on the effects of different antibiotics, namely timentin, cefotaxime, carbenicillin and ampicillin, on shoot regeneration of strawberry leaf explants showed the best shoot regeneration in the presence of 300 mg/L timentin and 150 mg/L cefotaxime. Assessment of the different factors affecting Agrobacterium mediated-transformation of strawberry with the luciferase gene showed the highest efficiency of putative transformant production (86%) in the treatment with no preculture, bacterial OD600 of 0.6 and the addition of 150 mg/L cefotaxime in the pre-selection and selection media. The presence of the luciferase gene in the plant genome was verified by the luciferase reporter gene assay, nested PCR amplification and dot blot of genomic DNA isolated from the young leaves of each putatively transformed plantlet.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Agriculture
Faculty of Biotechnology and Biomolecular Sciences
Institute of Tropical Agriculture
DOI Number: https://doi.org/10.3390/molecules20033647
Publisher: MDPI
Keywords: Stable transformation; Agrobacterium; Strawberry camarosa; PGreen0049 binary vector; Thidiazuron; Timentin; Cefotaxime
Depositing User: Mohd Hafiz Che Mahasan
Date Deposited: 08 Aug 2016 08:51
Last Modified: 08 Aug 2016 08:51
Altmetrics: http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.3390/molecules20033647
URI: http://psasir.upm.edu.my/id/eprint/43709
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